Antibodies against extracellular domains 2 and 3 or her2

Abstract

The present invention relates to an affinity ligand capable of selective interaction with a subset consisting of 37 consecutive amino acid residues or less from extracellular domains 2 and 3 of HER2, wherein the subset comprises the amino acid sequence LQVF and/or ESFDGD1 and to polypeptides consisting of such subsets.

1 .- 86 . (canceled) 
     
     
         87 . Affinity ligand capable of selective interaction with a subset consisting of 37 consecutive amino acid residues or less from extracellular domains 2 and 3 of Human Epidermal growth factor Receptor 2 (HER2) (SEQ ID NO:7), said subset comprising the amino acid sequence LQVF (SEQ ID NO:8) and/or ESFDGD (SEQ ID NO:9). 
     
     
         88 . Affinity ligand according to  claim 87 , wherein said subset consists of 26 amino acid residues or less. 
     
     
         89 . Affinity ligand according to  claim 88 , wherein said subset consists of an amino acid sequence selected from the group consisting of SEQ ID NO:11 and 15-20. 
     
     
         90 . Affinity ligand according to  claim 88 , wherein said subset consists of 21 amino acid residues or less. 
     
     
         91 . Affinity ligand according to  claim 87 , wherein said subset comprises LQVF (SEQ ID NO:8) and/or LPESFDGD (SEQ ID NO:11). 
     
     
         92 . Affinity ligand according to  claim 87 , wherein said subset consists of the sequence of amino acid residues 1-37 of SEQ ID NO:6. 
     
     
         93 . Affinity ligand according to  claim 87 , which inhibits growth of human breast cancer cells in culture by 20-100%. 
     
     
         94 . Affinity ligand according to  claim 93 , wherein the human breast cancer cells are BT474 breast cancer cells. 
     
     
         95 . Affinity ligand according to  claim 93 , which inhibits growth at a concentration of 500 ng/ml. 
     
     
         96 . Affinity ligand according to  claim 87 , which binds the subset with an EC50 of less than 100 nM. 
     
     
         97 . Affinity ligand according to  claim 87 , which is an antibody or a fragment or derivative thereof. 
     
     
         98 . Affinity ligand according to  claim 97 , which is a monoclonal antibody. 
     
     
         99 . Affinity ligand according to  claim 97 , which is a chimeric or humanized monoclonal antibody. 
     
     
         100 . A composition comprising an affinity ligand according to  claim 87  and a second affinity ligand capable of selective interaction with a second subset of 73 consecutive amino acid residues or less from extracellular domains 2 and 3 of Human Epidermal growth factor Receptor 2 (HER2) (SEQ ID NO:7), said second subset comprising one or more amino acid sequences selected from the group consisting of SEQ ID NO:12, SEQ ID NO:13 and SEQ ID NO:14. 
     
     
         101 . Composition comprising an affinity ligand according to  claim 87  and a tyrosine kinase inhibitor against Human Epidermal growth factor Receptor 2 (HER2). 
     
     
         102 . Method of preparation of a therapeutic antibody for the treatment of a disorder characterized by the overexpression of Human Epidermal growth factor Receptor 2 (HER2), comprising an immunization using a polypeptide which consists of 37 consecutive amino acid residues or less from extracellular domains 2 and 3 of HER2 (SEQ ID NO:7) and comprises the amino acid sequence LQVF (SEQ ID NO:8) and/or ESFDGD (SEQ ID NO:9) as an antigen. 
     
     
         103 . Method according to  claim 102 , wherein the therapeutic antibody is monoclonal. 
     
     
         104 . Method according to  claim 102 , wherein the therapeutic antibody is chimeric or humanized. 
     
     
         105 . Method of selection or purification of a therapeutic antibody for the treatment of a disorder characterized by the overexpression of Human Epidermal growth factor Receptor 2 (HER2), comprising the use of a polypeptide which consists of 37 consecutive amino acid residues or less from extracellular domains 2 and 3 of HER2 (SEQ ID NO:7) and comprises the amino acid sequence LQVF (SEQ ID NO:8) and/or ESFDGD (SEQ ID NO:9). 
     
     
         106 . Method for identification of an affinity ligand for treatment of a disorder characterized by the overexpression of Human Epidermal growth factor Receptor 2 (HER2), comprising the steps of:
 a) contacting a polypeptide comprising a subset according to  claim 87  with a putative affinity ligand in conditions that enable binding; and   b) determining whether the putative affinity ligand binds to the subset.   
     
     
         107 . Method according to  claim 106 , wherein the polypeptide of step a) consists of a subset consisting of 37 consecutive amino acid residues or less from extracellular domains 2 and 3 of HER2 (SEQ ID NO:7) and comprising the amino acid sequence LQVF (SEQ ID NO:8) and/or ESFDGD (SEQ ID NO:9). 
     
     
         108 . Method according to  claim 106 , wherein the disorder is a cancer, further comprising the step:
 c) determining whether the putative affinity ligand inhibits growth or induces apoptosis of cancer cells, such as BT474 breast cancer cells.   
     
     
         109 . Method of producing an affinity ligand, comprising:
 identifying an affinity ligand using the method according of  claim 106 ; and   producing said identified affinity ligand.   
     
     
         110 . Method for producing a clone comprising the steps of:
 a) providing cells obtained from a mammal which has been immunized with an antigen comprising a subset consisting of 37 consecutive amino acid residues or less from extracellular domains 2 and 3 of HER2 (SEQ ID NO:7) and comprising the amino acid sequence LQVF (SEQ ID NO:8) and/or ESFDGD (SEQ ID NO:9), which cells comprise DNA encoding an antibody capable of selective interaction with the subset; and   b) fusing said cells with myeloma cells to obtain at least one clone.   
     
     
         111 . Method according to  claim 110 , further comprising the step of:
 a′) immunizing the mammal with the antigen,   
       wherein step a′) precedes step a). 
     
     
         112 . Method according to  claim 110 , further comprising the step of:
 c) selecting a clone from step b) which expresses antibodies capable of selective interaction with the subset.   
     
     
         113 . Method according to  claim 110 , wherein the antigen of step a) consists of 37 consecutive amino acid residues or less from extracellular domains 2 and 3 of HER2 (SEQ ID NO:7) and comprises the amino acid sequence LQVF (SEQ ID NO:8) and/or ESFDGD (SEQ ID NO:9). 
     
     
         114 . Method according to  claim 110  or  112 , further comprising the step:
 d) providing a clone obtained in step b) or selected in step c), and merging 
 DNA from the clone, which DNA encodes at least the part of an antibody expressed by the clone that selectively interacts with the subset, with human antibody encoding DNA; and 
 e) incorporating the merged DNA from step d) in cells to obtain a clone for expression of a therapeutic antibody for treatment of a disorder characterized by the overexpression of Human Epidermal growth factor Receptor 2 (HER2). 
 
     
     
         115 . Method of producing an affinity ligand comprising: producing a clone using the method according to  claim 110 ; and obtaining said affinity ligand from said clone. 
     
     
         116 . Method of treatment of a mammalian subject having, or suspected of having, a disorder characterized by the overexpression of Human Epidermal growth factor Receptor 2 (HER2), comprising administering an effective amount of an affinity ligand according to  claim 87  or a composition comprising said affinity ligand and a second affinity ligand capable of selective interaction with a second subset of 73 consecutive amino acid residues or less from extracellular domains 2 and 3 of HER2 (SEQ ID NO:7), said second subset comprising one or more amino acid sequences selected from the group consisting of SEQ ID NO:12, SEQ ID NO:13 and SEQ ID NO:14, to the subject. 
     
     
         117 . Method according to  claim 116 , further comprising administering a tyrosine kinase inhibitor against HER2 to the subject. 
     
     
         118 . Method according to  claim 116 , wherein said subject has been treated by a therapeutic antibody capable of selective interaction with HER2, such as the extracellular domain of HER2, which therapeutic antibody is different from the affinity ligand. 
     
     
         119 . Method according to  claim 118 , wherein said disorder characterized by the overexpression of HER2 is a cancer and said cancer has developed resistance to the therapeutic antibody. 
     
     
         120 . Method according to  claim 116 , wherein the disorder characterized by the overexpression of HER2 is a cancer. 
     
     
         121 . Method according to  claim 120 , wherein the cancer is selected from the group consisting of breast cancer, squamos cell cancinoma, lung cancer, such as small cell or non-small cell lung cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, vulval cancer, liver cancer, hepatoma, colorectal cancer, such as colon cancer, endometrial carcinoma, salivary gland carcinoma, kidney cancer, thyroid cancer, Wilm's tumor, bladder cancer, endometrial cancer, renal cancer, head and neck cancer, gastric cancer, esophageal cancer and prostate cancer. 
     
     
         122 . Method according to  claim 121 , wherein the cancer is metastatic breast cancer. 
     
     
         123 . An article of manufacture, comprising a container, a composition within the container comprising an affinity ligand according to  claim 87  and a label on or associated with the container that indicates that said composition can be used for treating a disorder characterized by the overexpression of Human Epidermal growth factor Receptor 2 (HER2). 
     
     
         124 . Article of manufacture according to  claim 123 , wherein the container has a sterile access port. 
     
     
         125 . Article of manufacture according to  claim 123 , wherein the container is an intravenous solution bag or a vial having a stopper pierceable by a hypodermic needle. 
     
     
         126 . Article of manufacture according to  claim 123 , further comprising a second container comprising a pharmaceutically acceptable buffer.